Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

NO COMMERCIAL INTEREST

Hi Florian.  Could it be a fixation problem?  What is your fix solution?  Try
making your fix solutions up fresh immediately before the experiment.  This
helps avoid weird fixation artifacts.  Otherwise I would look to the antibody
as the culprit.  What is your source for the secondary?  Not all antibodies
are created equal: ergo, the reactivity can vary greatly between vendors.  We
find that Jackson Immunochemicals makes excellent secondary antibodies.

Good Luck!

Mike

>===== Original Message From Confocal Microscopy List
<[log in to unmask]> =====
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>--
>dear all,
>
>i am currently doing antibody stainings on paraffin sections.
>unfortunately, i am always getting very bright dots when i look at
>the staining under the confocal.when i do the control without first
>antibody, i also see these dots. therefore, they should clearly come
>from the 2nd antibody. it does not matter whether i use an alexa 488
>or alexa 546 coupled antibody, they both produce these effects. maybe
>the antibodies are bad, but they are from a commercial source and
>work fine in many applications. so, is this a probelm related to
>paraffin sectioning?
>
>i would appreciate any ideas or experience how to get around that
>problem and be forever grateful!
>
>
>best wishes,
>
>Florian
>-----------------------------------------------------------------------------
-----------------
>Florian Ulrich
>Heisenberg Lab
>Max-Planck-Institute for Molecular Cell Biology and Genetics
>Pfotenhauerstrasse 108
>01307 Dresden
>Germany
>phone: (+49) 351 210 2689
>fax:    (+49) 351 210 1489
>email: [log in to unmask]

>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>
Michael C. Adams
Microscopy & Imaging Manager for Dr. Clare Waterman-Storer
Laboratory of Cell Motility Studies
Department of Cell Biology
The Scripps Research Institute
Attn: Mail Code CB-163
10550 North Torrey Pines Road
La Jolla, CA 92037

TEL 858.784.9244
FAX 858.784.7521
EMAIL [log in to unmask]
>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>